Flow cytometric analyses of SAP expression Panel 1. Spleen cells from a C57BL/6 mouse were fixed and permeabilized using BD Cytofix™ Fixation Buffer (Cat. No. 554655) and BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were stained with APC Hamster Anti-Mouse CD3e (Cat. No. 553066) and either PE Rat IgG2a, κ Isotype Control (Cat. No. 553930; Left Plot) or PE Rat Anti-SAP antibody (Cat. No. 566729; Right Plot) at 0.125 μg/test. Two-color flow cytometric pseudocolor plots showing the correlated expression of SAP (or Ig Isotype control staining) versus CD3e were derived from gated events with the forward and side light-scatter characteristics of intact leucocytes. Panel 2. Human peripheral blood mononuclear cells (PBMCs) were similarly fixed and permeabilized and then stained with APC Mouse Anti-Human CD3 (Cat. No. 561811) and either PE Rat IgG2a, κ Isotype Control (Cat. No. 553930; Left Plot) or PE Rat Anti-SAP antibody (Right Plot) at 0.125 μg/test. Pseudocolor dot plots showing the correlated expression of SAP (or Ig Isotype control staining) versus CD3 were derived from gated events with the light-scatter characteristics of intact lymphocytes. Panel 3. Cells from the human Jurkat (Acute T Cell Leukemia, ATCC TIB-152) cell line were similarly fixed and permeabilized and then stained with PE Rat IgG2a, κ Isotype Control (dashed line histogram) or PE Rat Anti-SAP antibody (solid line histogram) at 0.125 μg/test. The fluorescence histogram showing SAP expression (or Ig Isotype control staining) was derived from gated events with the light-scatter characteristics of intact cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
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Flow cytometric analyses of SAP expression Panel 1. Spleen cells from a C57BL/6 mouse were fixed and permeabilized using BD Cytofix™ Fixation Buffer (Cat. No. 554655) and BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were stained with APC Hamster Anti-Mouse CD3e (Cat. No. 553066) and either PE Rat IgG2a, κ Isotype Control (Cat. No. 553930; Left Plot) or PE Rat Anti-SAP antibody (Cat. No. 566729; Right Plot) at 0.125 μg/test. Two-color flow cytometric pseudocolor plots showing the correlated expression of SAP (or Ig Isotype control staining) versus CD3e were derived from gated events with the forward and side light-scatter characteristics of intact leucocytes. Panel 2. Human peripheral blood mononuclear cells (PBMCs) were similarly fixed and permeabilized and then stained with APC Mouse Anti-Human CD3 (Cat. No. 561811) and either PE Rat IgG2a, κ Isotype Control (Cat. No. 553930; Left Plot) or PE Rat Anti-SAP antibody (Right Plot) at 0.125 μg/test. Pseudocolor dot plots showing the correlated expression of SAP (or Ig Isotype control staining) versus CD3 were derived from gated events with the light-scatter characteristics of intact lymphocytes. Panel 3. Cells from the human Jurkat (Acute T Cell Leukemia, ATCC TIB-152) cell line were similarly fixed and permeabilized and then stained with PE Rat IgG2a, κ Isotype Control (dashed line histogram) or PE Rat Anti-SAP antibody (solid line histogram) at 0.125 μg/test. The fluorescence histogram showing SAP expression (or Ig Isotype control staining) was derived from gated events with the light-scatter characteristics of intact cells. Flow cytometry and data analysis were performed using a BD LSRFortessa™ X-20 Cell Analyzer System and FlowJo™ software. Data shown on this Technical Data Sheet are not lot specific.
Product Details
BD Pharmingen™
SAP; Sh2d1a; SH2 domain-containing protein 1A
Mouse (QC Testing), Human (Tested in Development)
Rat IgG2a, κ
Mouse SAP Recombinant Protein
Intracellular staining (flow cytometry) (Routinely Tested)
0.2 mg/ml
AB_2869832
Aqueous buffered solution containing ≤0.09% sodium azide.
RUO
Preparation And Storage
Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. The antibody was conjugated with R-PE under optimum conditions, and unconjugated antibody and free PE were removed.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- An isotype control should be used at the same concentration as the antibody of interest.
- Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Species cross-reactivity detected in product development may not have been confirmed on every format and/or application.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
Stain Buffer (FBS) RUO
Size
500 mL
Cat No.
554656
Stain Buffer (BSA) RUO
Size
500 mL
Cat No.
554657
Fixation Buffer RUO
Size
100 mL
Cat No.
554655
Perm/Wash Buffer RUO
Size
100 mL
Cat No.
554723
PE Rat IgG2a, κ Isotype Control RUO
Size
0.1 mg
Cat No.
553930
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APC Hamster Anti-Mouse CD3e RUO
Size
0.1 mg
Cat No.
553066
566729 Rev. 1
Antibody Details
1A9
The 1A9 monoclonal antibody specifically recognizes Signaling lymphocytic activation molecule-associated protein (SLAM-associated protein or SAP). SAP is a cytoplasmic adaptor protein encoded by Sh2d1a (Src-Homology 2 Domain-Containing protein A1) that is expressed in T cells, NKT cells, NK cells, eosinophils, platelets, and some B cells. SAP contains an SH2 domain which can bind to immunoreceptor tyrosine-based switch motifs (ITSMs) present in the cytoplasmic domains of SLAM family receptors. This leads to the recruitment of various signaling molecules such as the Src-related protein tyrosine kinase Fyn and to the activation of downstream signaling pathways. Signaling through SLAM receptors enables cellular responses such as the production of various cytokines and activation of cytotoxicity. The 1A9 antibody crossreacts with human SAP which is encoded by the SH2D1A gene. Mutations in this gene are associated with some X-linked lymphoproliferative diseases. Dysregulation of the SLAM-SAP pathway might also play roles in some autoimmune diseases and nonprotective immune responses to infectious pathogens.
566729 Rev. 1
Format Details
PE
R-Phycoerythrin (PE), is part of the BD family of Phycobiliprotein dyes. This fluorochrome is a multimeric fluorescent phycobiliprotein with excitation maximum (Ex Max) of 496 nm and 566 nm and an emission maximum (Em Max) at 576 nm. PE is designed to be excited by the Blue (488 nm), Green (532 nm) and Yellow-Green (561 nm) lasers and detected using an optical filter centered near 575 nm (e.g., a 575/26-nm bandpass filter). As PE is excited by multiple lasers, this can result in cross-laser excitation and fluorescence spillover on instruments with various combinations of Blue, Green, and Yellow-Green lasers. Please ensure that your instrument’s configurations (lasers and optical filters) are appropriate for this dye.
PE
Yellow-Green 488 nm, 532 nm, 561 nm
496 nm, 566 nm
576 nm
566729 Rev.1
Citations & References
View product citations for antibody "566729" on CiteAb
Development References (5)
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Cannons JL, Tangye SG, Schwartzberg PL. SLAM family receptors and SAP adaptors in immunity. Annu Rev Immunol. 2011; 29:665-705. (Biology). View Reference
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Perez-Quintero LA, Roncagalli R, Guo H, Latour S, Davidson D, Veillette A. EAT-2, a SAP-like adaptor, controls NK cell activation through phospholipase Cgamma, Ca++, and Erk, leading to granule polarization. J Exp Med. 2014; 211(4):727-742. (Clone-specific: Western blot). View Reference
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Roncagalli R, Taylor JE, Zhang S, et al. Negative regulation of natural killer cell function by EAT-2, a SAP-related adaptor. Nat Immunol. 2005; 10(6):1002-1010. (Immunogen: Immunoprecipitation, Western blot). View Reference
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Wu N, Veillette A. SLAM family receptors in normal immunity and immune pathologies. Curr Opin Immunol. 2016; 38:45-51. (Biology). View Reference
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Zhong MC, Veillette A. Critical role of SAP in progression and reactivation but not maintenance of T cell-dependent humoral immunity. Mol Cell Biol. 2013; 33(6):1223-1232. (Clone-specific: Flow cytometry). View Reference
566729 Rev. 1
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.